title: Improvement in the visual discrimination of recombinant clones by size reduction of non-recombinant colonies.
abstract: A flexible approach circumventing cloning problems related to incomplete vector double digest is described. DNA methyltransferase gene insertion into MCS of commonly used expression vectors facilitates identification of both: i) the correct linear fragment in agarose gels due to the dilator effect, and ii) recombinant colonies by size and opacity differences resulting from methyltransferase toxicity.
authors.affiliation: Department of Microbiology, University of Gdansk, 80-308 Gdansk, Wita Stwosza 59, Poland. Electronic address: marian.sektas@biol.ug.edu.pl.
authors.forename: Marian
authors.initials: M
authors.lastname: Sektas
authors.forename: Beata
authors.initials: B
authors.lastname: Furmanek-Blaszk
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification; D005822:Genetic Vectors; D016254:Mutagenesis, Insertional
chemical_list: D004269:DNA, Bacterial; D004259:DNA-Directed DNA Polymerase
keywords: DNA methyltransferase; Positive/negative screening; Recombinant technology